Comparison of comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry and gas chromatography-mass spectrometry for the analysis of tobacco essential oils

A sample of tobacco essential oil was analyzed using gas chromatography-mass spectrometry (GUMS)and comprehensive two-dimensional gas chromatography coupled to a time-of-flight mass spectrometry (GC x GC/TOFMS), respectively. In the GUMS analysis, serially coupled columns were used. By comparing the GUMS results with GC x GC/TOFMS result,,, many more components in the essential oil could be found within the two-dimensional separation space of GC x GC. The quantitative determination of components in the essential oil was performed by GC x GC with flame ionization detection (FID), using a method of multiple internal standards calibration, (c) 2005 Elsevier B.V. All rights reserved.

Quantitative determination of compounds in tobacco essential oils by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry

A quantitative analysis of the individual compounds in tobacco essential oils is performed by comprehensive two-dimensional gas chromatography (GC x GC) combined with flame ionization detector (FID). A time-of-flight mass spectrometer (TOF/MS) was coupled to GC x GC for the identification of the resolved peaks. The response of a flame ionization detector to different compound classes was calibrated using multiple internal standards. In total, 172 compounds were identified with good match and 61 compounds with high probability value were reliably quantified. For comparative purposes, the essential oil sample was also quantified by one-dimensional gas chromatography-mass spectrometry (GC/MS) with multiple internal standards method. The results showed that there was close agreement between the two analysis methods when the peak purity and match quality in one-dimensional GC/MS are high enough. (c) 2005 Elsevier B.V. All rights reserved.

Analysis of the elements of catabolite repression in Clostridium acetobutylicum.

The ptsH gene, encoding the phosphotransferase protein HPr, from Clostridium acetobutylicum ATCC 824 was identified from the genome sequence, cloned and shown to complement a ptsH mutant of Escherichia coli. The deduced protein sequence shares significant homology with HPr proteins from other low-GC gram-positive bacteria, although the highly conserved sequence surrounding the Ser-46 phosphorylation site is not well preserved in the clostridial protein. Nevertheless, the HPr was phosphorylated in an ATP-dependent manner in cell-free extracts of C. acetobutylicum. Furthermore, purified His-tagged HPr from Bacillus subtilis was also a substrate for the clostridial HPr kinase/phosphorylase. This phosphorylation reaction is a key step in the mechanism of carbon catabolite repression proposed to operate in B. subtilis and other low-GC gram-positive bacteria. Putative genes encoding the HPr kinase/phosphorylase and the other element of this model, namely the catabolite control protein CcpA, were identified from the C. acetobutylicum genome sequence, suggesting that a similar mechanism of carbon catabolite repression may operate in this industrially important organism.

Agency and animation: the performance of interactive game characters

As an animator and practice-based researcher with a background in games development, I am interested in technological change in the video game medium, with a focus on the tools and technologies that drive game character animation and interactive story. In particular, I am concerned with the issue of ‘user agency’, or the ability of the end user to affect story development—a key quality of the gaming experience and essential to the aesthetics of gaming, which is defined in large measure by its interactive elements. In this paper I consider the unique qualities of the video game1 as an artistic medium and the impact that these qualities have on the production of animated virtual character performances. I discuss the somewhat oppositional nature of animated character performances found in games from recent years, which range from inactive to active—in other words, low to high agency. Where procedural techniques (based on coded rules of movement) are used to model dynamic character performances, the user has the ability to interactively affect characters in real-time within the larger sphere of the game. This game play creates a high degree of user agency. However, it lacks the aesthetic nuances of the more crafted sections of games: the short cut-scenes, or narrative interludes where entire acted performances are mapped onto game characters (often via performance capture)2 and constructed into relatively cinematic representations. While visually spectacular, cut-scenes involve minimal interactivity, so user agency is low. Contemporary games typically float between these two distinct methods of animation, from a focus on user agency and dynamically responsive animation to a focus on animated character performance in sections where the user is a passive participant. We tend to think of the majority of action in games as taking place via playable figures: an avatar or central character that represents a player. However, there is another realm of characters that also partake in actions ranging from significant to incidental: non-playable characters, or NPCs, which populate action sequences where game play takes place as well as cut scenes that unfold without much or any interaction on the part of the player. NPCs are the equivalent to supporting roles, bit characters, or extras in the world of cinema. Minor NPCs may simply be background characters or enemies to defeat, but many NPCs are crucial to the overall game story. It is my argument that, thus far, no game has successfully utilized the full potential of these characters to contribute toward development of interactive, high performance action. In particular, a type of NPC that I have identified as ‘pivotal’3—those constituting the supporting cast of a video game—are essential to the telling of a game story, particularly in genres that focus on story and characters: adventure games, action games, and role-playing games. A game story can be defined as the entirety of the narrative, told through non-interactive cut-scenes as well a interactive sections of play, and development of more complex stories in games clearly impacts the animation of NPCs. I argue that NPCs in games must be capable of acting with emotion throughout a game—in the cutscenes, which are tightly controlled, but also in sections of game play, where player agency can potentially alter the story in real-time. When the animated performance of NPCs and user agency are not continuous throughout the game, the implication is that game stories may be primarily told through short movies within games, making it more difficult to define video games animation as a distinct artistic medium.

Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana

Clarke, S. M., Mur, L. A. J., Wood, J. E., & Scott, I. M. (2004). Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana. The Plant Journal, 38(3), 432-447. Sponsorship: BBSRC RAE2008

The rpf gene of Micrococcus luteus encodes an essential secreted growth factor

Galina V. Mukamolova, Obolbek A. Turapov, Konstantin Kazarian, Miroslav Telkov, Arseny S. Kaprelyants, Douglas B. Kell and Michael Young (2002). The rpf gene of Micrococcus luteus encodes an essential secreted growth factor. Molecular Microbiology, 46 (3), 611-621. Sponsorship: BBSRC / Russian Foundation for Basic Research (grant 00-04-48691)/ WHO Global Programme for Vaccines and Immunization / Wellcome Trust RAE2008

Age- and activity-related differences in the abundance of Myosin essential and regulatory light chains in human muscle

Traditional methods for phenotyping skeletal muscle (e.g., immunohistochemistry) are labor-intensive and ill-suited to multixplex analysis, i.e., assays must be performed in a series. Addressing these concerns represents a largely unmet research need but more comprehensive parallel analysis of myofibrillar proteins could advance knowledge regarding age- and activity-dependent changes in human muscle. We report a label-free, semi-automated and time efficient LC-MS proteomic workflow for phenotyping the myofibrillar proteome. Application of this workflow in old and young as well as trained and untrained human skeletal muscle yielded several novel observations that were subsequently verified by multiple reaction monitoring (MRM).We report novel data demonstrating that human ageing is associated with lesser myosin light chain 1 content and greater myosin light chain 3 content, consistent with an age-related reduction in type II muscle fibers. We also disambiguate conflicting data regarding myosin regulatory light chain, revealing that age-related changes in this protein more closely reflect physical activity status than ageing per se. This finding reinforces the need to control for physical activity levels when investigating the natural process of ageing. Taken together, our data confirm and extend knowledge regarding age- and activity-related phenotypes. In addition, the MRM transitions described here provide a methodological platform that can be fine-tuned to suite multiple research needs and thus advance myofibrillar phenotyping.

Droga do ucznia. Polonistyczne ocenianie kształtujące na III i IV etapie edukacyjnym

Wydział Filologii Polskiej i Klasycznej: Instytut Filologii Polskiej

Herod Wielki i „nowa” Jerozolima

Summary: Herod the Great (73-4 B.C.E.) was a Roman client king of the small Jewish state Judaea in the last three decades before the common era. An essential aspect of Herod's reign was his role as a builder. Remarkably innovative, he created an astonishing record of architectural achievement, not only in Judaea but also throughout Greece and the Roman East. Herod’s own inclinations caused him to engage in a building program that paralleled that of his patron, Augustus. The most famous and ambitious project was the expansion of Jerusalem and rebuilding of the Second Temple. Josephus Flavius, a 1st-century Jewish historian, in his descriptions of the visual structure of Jerusalem delivers the picture of the Jewish society in the latter Second Temple Judaea, who were fundamentally antagonistic toward images. For Josephus, Roman iconography, such as Herod’s eagle from the Jerusalem Temple, represents not only political domination but also an unambiguous religious abomination. Visual conservatism in the public realm finds important verification in the excavated remains of Jerusalem’s Temple Mount and the Herodian Quarter (Upper City). Geometric patterns and forms predominate on the floor mosaic, stone furniture, in architectural detail and funerary remains. No human imagery is present in the Jewish context. However, Herodian structures in Jerusalem reflect the architectural and visual vocabulary of their time which contains popular elements of Roman domination in the ancient world.

Alterações metabólicas no diabético

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Distúrbios do metabolismo do cobre, ferro e zinco

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Investigation of lantibiotic regulation, immunity and synergy

Due to the increasing incidence of antibiotic resistant strains, the use of novel antimicrobials, such as bacteriocins, has become an ever more likely prospect. Lacticin 3147 (of which there are two components, Ltnα and Ltnβ) and nisin belong to the subgroup of bacteriocins called the lantibiotics, which has attracted much attention in recent years. The lantibiotics are antimicrobial peptides that contain unusual amino acids resulting from a series of enzyme-mediated post translational modifications. Given that there have been relatively few examples of lantibiotic-specific resistance; these antimicrobials appear to represent valid alternatives to classical antibiotics. However, the fact that lantibiotics are naturally only produced in small amounts often hinders their commercialisation. In order to overcome this bottleneck, several approaches can be employed. For example, we can create a situation that reduces the quantity of a lantibiotic required to inhibit a target by combining it with other antimicrobials. Here, following an initial screen involving lacticin 3147 and several classical antibiotics, it was observed between lacticin 3147 and the commercial antibiotics polymyxin B/E function synergistically. This reduced the amounts of the individual antimicrobials required for kill and broadened the spectrum of inhibition of both agents. Upon combination with polymyxins, lacticin 3147, which has been associated with Gram positive targets only, actively targeted Gram negative species such as Escherichia coli and Cronobacter sp. An alternative means of addressing problems associated with lantibiotic yield is to better understand how production is regulated, and ultimately use this information to enhance peptide levels. With this in mind the regulation of lacticin 3147 production from the promoter Pbac was investigated using a green fluorescent protein (GFP) expression reporter system. This revealed that elements within both of the divergent operons of the lacticin 3147 gene cluster are involved in Pbac regulation. That is, LtnR, already established as a negative regulator of itself and the lacticin 3147 associated immunity genes, also acts as an activator of Pbac transcription. In contrast, an enhanced level of expression is observed in the absence of the lacticin 3147 structural genes, ltnA1 and ltnA2, indicating that these genes/gene products are involved in Pbac repression. In fact, through complementation of the ltnA2 gene, it was revealed that this regulation is more likely to be dependent on the presence of the gene transcript rather that the corresponding prepropeptide or modified Ltnβ. It may be that if lacticin 3147 production is successfully enhanced, the ability of the producing cell to protect itself may become an issue. To prepare for such a possibility a bioengineered derivative of the lacticin 3147 immunity protein LtnI (LtnI I81V) which provides enhanced protection was discovered through an in depth investigation involving the site and saturation mutagenesis of this protein. In addition, the creation of truncated forms of LtnI allowed the identification of important and essential regions of this immunity protein. Finally, as mentioned, self-immunity is essential to prevent self-killing. However the discovery of nisin U immunity and regulatory gene homologues (spiFEGRR’K) within the pathogenic strain S. infantarius subsp. infantarius is a cause for concern as it represents an example of immune mimicry, a form of lantibiotic-specific resistance. The ability of spiFEG to confer protection was apparent when they successfully provided protection to nisin A, F, Z, Q and U when expressed heterologously in the nisin sensitive L. lactis HP host. As a consequence of the studies presented in this thesis, it is likely that strategies will emerge that will facilitate the production of greater levels of lacticin 3147 production and lead to enhanced immunity in lactococcal backgrounds. Alternatively the need for enhanced production could be avoided through the use of antimicrobial combinations. In addition, providing awareness of the threats of the emergence of resistance through immune mimicry can allow researchers to develop strategies to prevent this phenomenon from leading to the dissemination of lantibiotic resistance.

Biomethane production from grass silage: laboratory assessment to maximise yields

On-farm biogas production is typically associated with forage maize as the biomass source. Digesters are designed and operated with the focus of optimising the conditions for this feedstock. Thus, such systems may not be ideally suited to the digestion of grass. Ireland has ca. 3.85 million ha of grassland. Annual excess grass, surplus to livestock requirements, could potentially fuel an anaerobic digestion industry. Biomethane associated with biomass from 1.1 % of grassland in Ireland, could potentially generate over 10 % renewable energy supply in transport. This study aims to identify and optimise technologies for the production of biomethane from grass silage. Mono-digestion of grass silage and co-digestion with slurry, as would occur on Irish farms, is investigated in laboratory trials. Grass silage was shown to have 7 times greater methane potential than dairy slurry on a fresh weight basis (107 m3 t-1 v 16 m3 t-1). However, comprehensive trace element profiles indicated that cobalt, iron and nickel are deficient in mono-digestion of grass silage at a high organic loading rate (OLR) of 4.0 kg VS m-3 d-1. The addition of a slurry co-substrate was beneficial due to its wealth of essential trace elements. To stimulate hydrolysis of high lignocellulose grass silage, particle size reduction (physical) and rumen fluid addition (biological) were investigated. In a continuous trial, digestion of grass silage of <1 cm particle size achieved a specific methane yield of 371 L CH4 kg-1 VS when coupled with rumen fluid addition. The concept of demand driven biogas was also examined in a two-phase digestion system (leaching with UASB). When demand for electricity is low it is recommended to disconnect the UASB from the system and recirculate rumen fluid to increase volatile fatty acid (VFA) and soluble chemical oxygen demand (SCOD) production whilst minimising volatile solids (VS) destruction. At times of high demand for electricity, connection of the UASB increases the destruction of volatiles and associated biogas production. The above experiments are intended to assess a range of biogas production options from grass silage with a specific focus on maximising methane yields and provide a guideline for feasible design and operation of on-farm digesters in Ireland.

Religious memory as a conveyor of authoritative tradition: The necessary and essential component in a definition of religion

In a landmark book published in 2000, the sociologist Danièle Hervieu-Léger defined religion as a chain of memory, by which she meant that within religious communities remembered traditions are transmitted with an overpowering authority from generation to generation. After analysing Hervieu-Léger’s sociological approach as overcoming the dichotomy between substantive and functional definitions, this article compares a ritual honouring the ancestors in which a medium becomes possessed by the senior elder’s ancestor spirit among the Shona of Zimbabwe with a cleansing ritual performed by a Celtic shaman in New Hampshire, USA. In both instances, despite different social and historical contexts, appeals are made to an authoritative tradition to legitimize the rituals performed. This lends support to the claim that the authoritative transmission of a remembered tradition, by exercising an overwhelming power over communities, even if the memory of such a tradition is merely postulated, identifies the necessary and essential component for any activity to be labelled “religious”.

Sorting under partial information (without the ellipsoid algorithm)

We revisit the well-known problem of sorting under partial information: sort a finite set given the outcomes of comparisons between some pairs of elements. The input is a partially ordered set P, and solving the problem amounts to discovering an unknown linear extension of P, using pairwise comparisons. The information-theoretic lower bound on the number of comparisons needed in the worst case is log e(P), the binary logarithm of the number of linear extensions of P. In a breakthrough paper, Jeff Kahn and Jeong Han Kim (STOC 1992) showed that there exists a polynomial-time algorithm for the problem achieving this bound up to a constant factor. Their algorithm invokes the ellipsoid algorithm at each iteration for determining the next comparison, making it impractical. We develop efficient algorithms for sorting under partial information. Like Kahn and Kim, our approach relies on graph entropy. However, our algorithms differ in essential ways from theirs. Rather than resorting to convex programming for computing the entropy, we approximate the entropy, or make sure it is computed only once in a restricted class of graphs, permitting the use of a simpler algorithm. Specifically, we present: an O(n2) algorithm performing O(log n·log e(P)) comparisons; an O(n2.5) algorithm performing at most (1+ε) log e(P) + Oε(n) comparisons; an O(n2.5) algorithm performing O(log e(P)) comparisons. All our algorithms are simple to implement. © 2010 ACM.